Duque-Guimarães Daniella E, de Almeida-Faria Juliana, Ong Thomas Prates, Ozanne Susan E
University of Cambridge Metabolic Research Laboratories and MRC Metabolic Diseases Unit, Wellcome Trust-MRC Institute of Metabolic Science, Addenbrooke's Hospital, Cambridge, UK.
Department of Physiology and Biophysics, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil.
Methods Mol Biol. 2017;1546:149-159. doi: 10.1007/978-1-4939-6730-8_11.
Pulsed stable isotope labeling by amino acids in cell culture (pSILAC) comprises a variation of the classical SILAC proteomic methodology that enables the identification of short-term proteomic responses such as those elicited by micro RNAs (miRNAs). Here, we describe a detailed pSILAC protocol for global identification and quantification of protein translation alterations induced by a miRNA using 3T3-L1 pre-adipocytes as a model system.
细胞培养中氨基酸脉冲稳定同位素标记（pSILAC）是经典SILAC蛋白质组学方法的一种变体，可用于识别短期蛋白质组反应，如由微小RNA（miRNA）引发的反应。在此，我们描述了一种详细的pSILAC方案，以3T3-L1前脂肪细胞为模型系统，用于全面鉴定和定量由miRNA诱导的蛋白质翻译变化。
